Leukemias, lymphomas, carcinomas and other malignancies are well known and described in, e.g., Harrison's Principles of Internal Medicine, Wilson et al., eds., McGraw-Hill, New York, pp. 1599-1612. These malignancies appear to have somehow escaped the immune system surveillance mechanisms that eliminate rapidly and continuously proliferating cells. The exact mechanism by which these malignancies escape the immune system surveillance is not known.
Some of these malignant immune system cells are malignant antigen presenting cells which do not function properly within the immune cascade. For example, neoplastic B cells cannot induce even weak allogeneic or autologous mixed lymphocyte reactions in vitro. Further evidence that malignancies survive due to the failure of the immune surveillance mechanism includes the increased frequency of such malignancies in immunocompromised individuals, such as allograft recipients and those receiving long-term immunosuppressant therapy. Further, the frequency of these malignancies is increased in patients having Acquired Immune Deficiency Syndrome (AIDS) and patients with primary immune deficiency syndromes, such as X-linked lymphoproliferative syndrome or Wiscott-Aldrich Syndrome (Thomas et al., Adv. Cancer Res. 57:329, 1991).
The immune system normally functions to eliminate malignant cells by recognizing the malignant cells as foreign cells and clearing those cells from the body. An immune reaction depends on both the immune system's antibody response and on the cellular immune response within a patient. More specifically, the cellular immune response which acts to recognize the malignant cells as foreign requires a number of different cells of the immune system and the interaction between those cells. An immune reaction begins with a T lymphocyte (T cell) which has on its cell surface the T cell receptor. The T cell also has the ability to express on its surface various accessory molecules which interact with accessory molecules on the B lymphocyte (B cell). When the T cell receptor of the T cell specifically binds to a foreign antigen, such as a malignant cell, it becomes activated and expresses the accessory molecule ligand, CD40 ligand on its cell surface. The accessory cell molecule ligand is only present on the activated T cells for a short period of time and is rapidly removed from the cell surface. After the accessory cells molecule ligand is removed from the surface of the activated T cell, its ability to bind to B cells via the accessory molecule ligand is destroyed.
When present on the surface of an activated T cell, the accessory cell ligand can specifically bind to the accessory cell molecule present on the B cell. This specific T-B cell interaction causes the B and T cell to express costimulutory surface accessory molecule and cytokines which result in an immune activation which lead to cytolytic T cells which specifically kill and remove the malignant cell from the body.
The interaction with an activated T cell is not solely limited to B cells but rather can be carried out by any cell which is able to present antigen to the T cell (an antigen presenting cell). These cells include B lymphocyte, macrophages, dendritic cells, monocytes, Langerhans cells, interdigitating cells, follicular dendritic cells or Kupffer cells. These cells all are known to have various accessory molecules on the cell surface which allow them to interact with other cells of the immune system. For example, these antigen presenting cells all have the accessory molecule CD40 on their cell surface. The presence of these accessory molecules allows these antigen presenting cells to specifically bind to complimentary accessory molecule ligand and thus directly interact with other immune cells.
A large number of accessory molecule ligands are members of the tumor necrosis factor superfamily. (Fanslow et al., Sem. Immun., 6:267-268 (1994). The genes for a number of these accessory molecule ligands have been cloned and identified. These accessory molecule ligand genes encode accessory molecules which all have the configuration of Type II membrane proteins and exhibit varying degrees of homology with other accessory molecule ligand genes. For example, the accessory molecule ligand genes encoding both murine CD40 ligand and human CD40 ligand have been isolated. See, Armitage et al., Nature, 357:80-82 (1992) and Hollenbaugh et al., EMBO J., 11:4313-4321 (1992).
CD40 and its ligand, CD40 ligand are critical components of a normal immune response. CD40 mediated signals induce immune lymphocytes to proliferate and differentiate and become potent antigen presenting cells. Malignant or neoplastic B cells are poor antigen presenter cells and are unable to stimulate a vigorous allogeneic mixed lymphocyte reaction. Successful cross linking of CD40 molecules on immune cells results in a strong allogeneic mixed lymphocyte reaction suggesting a strong immune reaction. Various soluble CD40 ligands or antibodies specific for CD40 have been used to potentially cross link CD40. These soluble CD40 ligands and CD40-specific antibodies are not optimal for cross linking the CD40 molecules on antigen presenting cells and do not work as effectively as CD40 ligand expressed on a cell membrane to produce strong stimulation of antigen presenting cells. These methods are also difficult to implement because large amounts of CD40 ligand constructs or antibodies must be isolated which is difficult and time-consuming work. Other strategies to utilize CD40 ligand in solution or as a membrane bound molecule including transformation of fibroblasts with CD40 ligand to produce cultured cells which are then used to present antigen are not amenable to in vivo human clinical protocols.
CD95 (Fas) interaction with its ligand (Fas-ligand, or FasL) functions to limit the duration of the immune response and/or life-span of activated lymphocytes. Apoptosis induced by Fas-FasL binding serves to clear activated self-reactive lymphocytes. Problems caused by altering this pathway have been demonstrated in animals with defects in Fas<->Fas-ligand interactions. Mice having mutations, which inactivate CD95 or FasL, develop numerous disorders including autoimmune pathology resembling that seen in patients with rheumatoid arthritis (RA) or systemic lupus. Zhang, et al., in J. Clin. Invest. 100:1951-1957 (1997) show that injection of FasL-expressing virus, into the joints of mice with collagen-induced-arthritis, results in apoptosis of synovial cells and relief of arthritis symptoms. Expression of Fas ligand allows clearance of activated cells which play a role in the pathogenesis of autoimmune disease. Therefore, a gene therapy strategy for introducing FasL into the joints of rheumatoid arthritis patients could function to improve disease pathology by leading to destruction of the infiltrating mononuclear cells.
Administration of soluble accessory molecules and accessory molecule ligands has been shown to trigger or to be associated with adverse physiological effects. For example, treatment of mice, having wild-type CD40-receptor expression, with soluble CD40L-CD8 fusion protein resulted in a pulmonary inflammatory response. This was not observed in mice in which the gene for the CD40 receptor had been knocked out. These experiments, described in Wiley, J. A. et al., Journal of Immunology 158:2932-2938 (1997), support in vitro data which suggest that CD40 ligation can result in inflammatory responses.
Direct administration of purified recombinant soluble Tumor Necrosis Factor (either α or β) results in shock and tissue injury, as. described in Tracey, K. J., and A. Cerami, Annu. Rev. Med. 45:491-503 (1994). Within minutes after acute intravenous or intra-arterial administration of TNF, a syndrome of shock, tissue injury, capillary leakage syndrome, hypoxia, pulmonary edema, and multiple organ failure associated with a high mortality ensues. Chronic low dose of TNF causes anorexia, weight loss, dehydration and depletion of whole-body protein and lipid.
Soluble Fas ligand and receptor have also been shown to be associated with tissue damage and other adverse effects. CD95, the Fas receptor, is a mediator of apoptosis. Fas ligand induces apoptosis by binding to Fas receptor. As shown in Galle, P. R., et al., J. Exp. Med. 182:1223-1230 (1995) administering an agonistic anti-Fas antibody resulted in liver damage to mice. Mice injected intraperitoneally with the agonistic antibody died within several hours, and analyses revealed that severe liver damage by apoptosis was the most likely cause of death.
The role of soluble Fas ligand (FasL), in the pathogenesis of systemic tissue injury in aggressive lymphoma is described in Sato, K. et al., British Journal of Haematology, 94:379-382 (1996). The findings presented in this report indicate that soluble FasL is directly associated with the pathogenesis of liver injury and pancytopenia.
CD27, the receptor for the accessory molecule ligand, CD70, was shown, in a report written by van Oers, et al., in Blood 82:3430-3436 (1993), to be associated with B cell malignancies.
The above findings all contraindicate the administration of soluble accessory molecule ligands, highlighting the need for therapies that increase the levels of these molecules without resulting in an elevation of their soluble forms.
Despite the wealth of information regarding accessory molecule ligand genes and their expression on the surface of various immune cells, the exact mechanism by which the accessory molecule ligand genes are regulated on antigen presenting cells is not yet known. Without specific knowledge of the regulation of expression of accessory molecule ligand genes on these antigen presenting cells, altering the immune response by varying expression of an accessory molecule ligand gene has to date not been possible. Without any specific knowledge as to how to regulate the expression of an accessory molecule ligand gene on an antigen presenting cell, it is not possible to alter the immune response towards malignant cells. Thus, there was a need for a method of increasing the expression of an accessory molecule ligand gene on normal and malignant cells including antigen presenting cells.
Further, without the ability to regulate the expression of accessory molecule ligands, it is not possible to alter the immune clearance of these cells.